1.Studies on distribution and imaging of new 99Tcm-labelled RGD peptide in synovial pannus of mice
Gula DA ; Hongbin LI ; Yong WANG
Chinese Journal of Rheumatology 2013;17(10):661-666,后插2
Objective To observe the distribution and imaging of radiolabelled arginine-glycineaspartic(RGD)peptide(99Tcm-3P4-RGD2)in collagen-induced arthritis(CIA),and to investigate the possibility of new markers for monitoring the angiogenesis of rheumatoid arthritis(RA)before and after treatment by single photon emission computed tomography(SPECT)labelled with 99Tcm-3P4-RGD2.Methods Type Ⅱ chicken collagen was injected into the tails of Wistar rats to establish arthritis models.The CIA-Wistar rats were randomly divided into three group:the Avastin group,the recombinant human tumor necrosis factor-α receptor Ⅱ group,and the blank control group.The changes of the arthritis index,joint pad thickness and radiological of T/NT before and after treatment were observed.The level of vascular endothelial growth factor(VEGF)in the serum was measured by enzyme-linked immuno sorbent assay(ELISA).The back metapedes pathological section was analyzed by Hematoxylin and Eosin stain(HE)and Safranin O stain as well as arthritic pathology index was also scored.The synovial expression level of VEGF was measured by immuno-histochemistry.The stain level of VEGF was assayed by semiquantitative approach.T test was applied for the comparison of two intergroup samples.Results Abnormal radioactive concentration appeared in the arthritis of the CIA-Wistar rats(1.40±0.17,1.32±0.20,1.30±0.08,P<0.05).Through the treatment of Avastin and recombinant human tumor necrosis factor-α receptor Ⅱ,the radiological of T/NT decreased significantly(0.43±0.14,0.40±0.12,t=17.710,16.812,P<0.05).After the treatment with Avastin and recombinant human tumor necrosis factor-α receptor Ⅱ,the severity of synovial pannus,joint pad thickness,arthritis index and the expression of VEGF were all decreased.The radiological of T/NT was positively correlated with the indices mentioned above(r=0.753,0.800,0.892,all P<0.01 respectively with the the rhTNFR:Fc group)(r=0.701,P=0.001;r=0.502,P=0.024;r=0.481,P=0.032 individually with the rhTNFR:Fc group).Conclusion When 99Tcm-3P4-RGD2 is injected through the tail vein of Wistar rats,it can show the pathological changes of arthritis clearly.99Tcm-3P4-RGD2may be the new eikonogen to monitor the angiogenesis of RA before and after treatment.
2.Experimental study of miR-7 antagomir in treatment of MRL/lpr mice
Gula DA ; Jing WANG ; Yong WANG ; Hongbin LI
Chinese Journal of Immunology 2024;40(4):680-685,692
Objective:To investigate the efficacy of miR-7 antagomir in treatment of MRL/lpr mice.Methods:Thirty MRL/lpr mice were selected and randomly divided into miR-7 antagomir group,CTX group and Saline control group,with 10 mice in each group.MRL/lpr mice were treated at 13 weeks,and the above drugs were administered continuously for 5 weeks.General condition of the mice in each group was observed weekly and weighed,and serum and urine samples were collected.Serum ANA,anti-ds-DNA an-tibody and C3 levels of mice in the 13~17 weeks of three groups were detected by ELISA,and urinary protein level of mice in the above three groups were detected at every week.Changes in proportion of Tfh cells in the three groups of mice were detected by flow cytometry.RT-PCR was used to detect expression levels of miR-7 and PTEN mRNA in spleen and lymph nodes of three groups of mice.Pathological conditions of kidneys of three groups of mice were observed under light microscope by HE,PAS,PASM staining.Results:After 5 weeks of treatment,in terms of gross morphology:Compared with Saline group,mice in miR-7 antagomir group and CTX group were in good general condition,and their body weight were increased with the increased age.In terms of serology:Serum ANA and anti-ds-DNA antibody of miR-7 antagomir group and CTX group were significantly lower than those of Saline group,while level of complement C3 was significantly increased,there was no statistical difference in the above indicators between the two treatment groups.In terms of cytology:Proportion of Tfh cells in spleen of mice in miR-7 antagomir group was significantly lower than that in Saline group.Compared with Saline group,expression level of miR-7 in spleen B cells of mice in miR-7 antagomir group was decreased,while expression level of PTEN mRNA was increased.In addition,pathological changes of kidneys of mice in miR-7 antagomir group and CTX group were improved compared with Saline group,there was no significant difference in pathological changes of kidneys between the two groups.Conclusion:miR-7 can be used as a potential target for treatment of systemic lupus erythematosus,miR-7 antagomir can show similar therapeutic effects as the classical immunosuppressant CTX,effectively improve the lupus-like disease manifestations in MRL/lpr mice and delay the progression of the disease.